OBJECTIVES: To investigate the expression of CDKN3 in gastric cancer and its impact on prognosis of gastric cancer patients. METHODS: We analyzed CDKN3 expression in clinical specimens from 114 gastric cancer patients and assessed its association with 5-year postoperative survival of the patients. GO and KEGG enrichment analyses were used to predict the biological function and possible mechanism of CDKN3. The effects of lentivirus-mediated CDKN3 knockdown on biological behaviors of gastric cancer cells were evaluated using Transwell assay, CCK-8 assay, TUNEL staining, flow cytometry, and Western blotting. RESULTS: CDKN3 expression was significantly higher in gastric cancer tissues than in the adjacent tissues with significant correlations with CEA level, CA19-9 level, and T and N staging (P<0.05). High CDKN3 expression was an independent risk factor affecting 5-year postoperative survival of the patients and predictive for long-term prognosis (P<0.01). Enrichment analyses suggested a probable association of CDKN3 with apoptosis. In MGC-803 cells, CDKN3 knockdown significantly lowered migration and invasion capacities of the cells, while CDKN3 overexpression produced the opposite effects. TUNEL staining revealed a significantly lower level of cell apoptosis in gastric cancer tissues than in adjacent tissues (P<0.01). CDKN3 knockdown obviously inhibited proliferation and increased apoptosis of MGC-803 cells. CDKN3 overexpression down-regulated the expressions of p53, p21 and Bax and up-regulated the expressions of p-p65 and Bcl-2. CONCLUSIONS CDKN3 is highly expressed in gastric cancer tissues and affects patient prognosis. CDKN3 overexpression promotes proliferation, invasion and migration and suppressed apoptosis of gastric cancer cells possibly through the p53/NF-κB signaling pathway.
Humans ; Stomach Neoplasms/pathology* ; Apoptosis ; Signal Transduction ; Tumor Suppressor Protein p53/metabolism* ; Cell Movement ; Cell Line, Tumor ; NF-kappa B/metabolism* ; Prognosis ; Cyclin-Dependent Kinase Inhibitor Proteins/metabolism* ; Cell Proliferation ; Neoplasm Invasiveness ; Male ; Female ; Dual-Specificity Phosphatases

Objective:To investigate the influencing factors and prognosis of early recurrence after gastrectomy for gastric cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 2 078 patients who underwent gastrectomy for gastric cancer at six medical centers across China, including Fudan University Shanghai Cancer Center et al, between January 2012 and June 2023 were collected. There were 1 449 males and 629 females, aged (59±11) years. Patients were classified as early recurrence and late recurrence based on the time of post-operative recurrence. Observation indicators: (1) comparison of clinicopathological characteristics between gastric cancer patients with different recurrence types; (2) recurrence and metastasis of tumor; (3) survival of patients after postoperative recurrence of gastric cancer; (4) analysis of influencing factors for early recurrence after gastrectomy for gastric cancer. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the rank sum test. Multivariate analysis was conducted using the Logistic regression model. Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. Results:(1) Comparison of clinicopathological characteristics between gastric cancer patients with different recurrence types. Among the 2 078 patients, 1 452 cases had early recurrence and 626 cases had late recurrence. There were significant differences in preoperative carcinoembryonic antigen, preoperative CA19-9, preoperative CA72-4, preoperative albumin, tumor diameter, neoadjuvant therapy, R 0 resection, combined organ resection, scope of gastric resection, nerve and vessel infiltration, degree of tumor differentiation, pathological N staging, pathological TNM staging between early and late recurrence patients ( P<0.05). (2) Recurrence and metastasis of tumor. Among the 2 078 patients, 200 cases had local recurrence, 1 213 cases had hematogenous metastases, 392 cases had distant lymph node metastases, and 731 cases had peritoneal metastases. Among the 1 452 early recurrence patients, 142 cases had local recurrence, 834 cases had hematogenous metastases, 289 cases had distant lymph node metastases, and 507 cases had peritoneal metastases. Among the 626 late recurrence patients, 58 cases had local recurrence, 379 cases had hematogenous metastases, 103 cases had distant lymph node metastases, and 224 cases had peritoneal metastases. One patient may have multiple forms of recurrence and metastasis. There was no significant difference in the above indica-tors between early and late recurrence patients ( χ2=0.13, 1.74, 3.40, 0.14, P>0.05). (3) Survival of patients after postoperative recurrence of gastric cancer. All 2 078 patients were followed up until death after recurrence, with a follow-up time of 31(range, 9?147)months. The 1-, 2-, 3-, and 5-year overall survival rates after recurrence were 33.5%, 17.2%, 10.1%, and 3.3% in early recurrence patients, versus 44.2%, 21.6%, 12.8%, and 5.8% in late recurrence patients, respectively, showing a significant difference in overall survival after recurrence between the two groups ( hazard ratio=0.84, 95% confidence interval as 0.76?0.92, P<0.05). (4) Analysis of influencing factors for early recurrence after gastrectomy for gastric cancer. Results of multivariate analysis showed that combined organ resection, total gastrectomy, pathological TNM staging as stage Ⅲ were independent risk factors for early recurrence after gastrectomy for gastric cancer ( odds ratio=1.31, 1.32, 1.34, 95% confidence interval as 1.01?1.70, 1.06?1.65, 1.05?1.71, P<0.05) and normal preoperative tumor markers, neoadjuvant therapy, R 0 resection were independent protective factors for early recurrence ( odds ratio=0.61, 0.50, 0.38, 95% confidence interval as 0.49?0.76, 0.35?0.72, 0.25?0.58, P<0.05). Conclusions:Compared with patients with late recurrence after gastric cancer surgery, patients with early recurrence have a poor prognosis, in which liver metastases is more common. Combine organ resection, total gastrectomy, pathological TNM staging as stage Ⅲ are independent risk factors for early recurrence, and normal preoperative tumor markers, neoadjuvant therapy, R 0 resection are independent protective factors for early recurrence after gastrectomy for gastric cancer.

Objective:To investigate the influencing factors and prognosis of early recurrence after gastrectomy for gastric cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 2 078 patients who underwent gastrectomy for gastric cancer at six medical centers across China, including Fudan University Shanghai Cancer Center et al, between January 2012 and June 2023 were collected. There were 1 449 males and 629 females, aged (59±11) years. Patients were classified as early recurrence and late recurrence based on the time of post-operative recurrence. Observation indicators: (1) comparison of clinicopathological characteristics between gastric cancer patients with different recurrence types; (2) recurrence and metastasis of tumor; (3) survival of patients after postoperative recurrence of gastric cancer; (4) analysis of influencing factors for early recurrence after gastrectomy for gastric cancer. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the rank sum test. Multivariate analysis was conducted using the Logistic regression model. Kaplan-Meier method was used to calculate survival rate and plot survival curve, and Log-rank test was used for survival analysis. Results:(1) Comparison of clinicopathological characteristics between gastric cancer patients with different recurrence types. Among the 2 078 patients, 1 452 cases had early recurrence and 626 cases had late recurrence. There were significant differences in preoperative carcinoembryonic antigen, preoperative CA19-9, preoperative CA72-4, preoperative albumin, tumor diameter, neoadjuvant therapy, R 0 resection, combined organ resection, scope of gastric resection, nerve and vessel infiltration, degree of tumor differentiation, pathological N staging, pathological TNM staging between early and late recurrence patients ( P<0.05). (2) Recurrence and metastasis of tumor. Among the 2 078 patients, 200 cases had local recurrence, 1 213 cases had hematogenous metastases, 392 cases had distant lymph node metastases, and 731 cases had peritoneal metastases. Among the 1 452 early recurrence patients, 142 cases had local recurrence, 834 cases had hematogenous metastases, 289 cases had distant lymph node metastases, and 507 cases had peritoneal metastases. Among the 626 late recurrence patients, 58 cases had local recurrence, 379 cases had hematogenous metastases, 103 cases had distant lymph node metastases, and 224 cases had peritoneal metastases. One patient may have multiple forms of recurrence and metastasis. There was no significant difference in the above indica-tors between early and late recurrence patients ( χ2=0.13, 1.74, 3.40, 0.14, P>0.05). (3) Survival of patients after postoperative recurrence of gastric cancer. All 2 078 patients were followed up until death after recurrence, with a follow-up time of 31(range, 9?147)months. The 1-, 2-, 3-, and 5-year overall survival rates after recurrence were 33.5%, 17.2%, 10.1%, and 3.3% in early recurrence patients, versus 44.2%, 21.6%, 12.8%, and 5.8% in late recurrence patients, respectively, showing a significant difference in overall survival after recurrence between the two groups ( hazard ratio=0.84, 95% confidence interval as 0.76?0.92, P<0.05). (4) Analysis of influencing factors for early recurrence after gastrectomy for gastric cancer. Results of multivariate analysis showed that combined organ resection, total gastrectomy, pathological TNM staging as stage Ⅲ were independent risk factors for early recurrence after gastrectomy for gastric cancer ( odds ratio=1.31, 1.32, 1.34, 95% confidence interval as 1.01?1.70, 1.06?1.65, 1.05?1.71, P<0.05) and normal preoperative tumor markers, neoadjuvant therapy, R 0 resection were independent protective factors for early recurrence ( odds ratio=0.61, 0.50, 0.38, 95% confidence interval as 0.49?0.76, 0.35?0.72, 0.25?0.58, P<0.05). Conclusions:Compared with patients with late recurrence after gastric cancer surgery, patients with early recurrence have a poor prognosis, in which liver metastases is more common. Combine organ resection, total gastrectomy, pathological TNM staging as stage Ⅲ are independent risk factors for early recurrence, and normal preoperative tumor markers, neoadjuvant therapy, R 0 resection are independent protective factors for early recurrence after gastrectomy for gastric cancer.

Objective To investigate the apoptotic effect of petasin on myeloma RPMI 8226 cells and the mechanisms.Methods The inhibition of petasin on the proliferation of myeloma RPMI 8226 cells was tested by trypan blue assay.Apoptosis of RPMI 8226 cells was measured by terminal-deoxynueleotidyl transferase mediated dUTP nick end labeling (TUNEL)assay and Hoechst 33258 staining assay.Effects of petasin on caspase-3,8 and 9 expressions,phosphorylation of ERK1/2,MEK(p-ERK1/2 ;p-MEK)and p38MAPK(p-p38MAPK)protein were analyzed by Western blot.Results Incubation by petasin for 24 h,48 h or 72 h could significantly inhibit the pro-liferation of myeloma RPMI 8226 cells (P <0.01,P <0.01,P <0.01).Petasin induced the apoptosis of myeloma RPMI 8226 cells in time-and concentration-dependent manners (P <0.05,P <0.05).Caspase inhibitor pretreat-ment could significantly inhibit the apoptosis of myeloma cells.After cultured with petasin for 72 h,the expressions of caspase-3,8 and 9 were obviously enhanced (P <0.05,P <0.01,P <0.05)and phosphorylation of p-p38MAPK of RPMI8226 cells was significantly increased (P <0.01).However,phosphorylation of p-ERK1/2 and p-MEK was decreased significantly (P <0.01,P <0.05).Conclusion Petasin can inhibit the proliferation of myeloma RPMI 8226 cells and induce apoptosis.The mechanism may be related to the activation of caspase-3,8 and 9 proteins and the changes in phosphorylation of p38MAPK,ERK1/2 and MEK.

Objective To investigate the expression of aquaporin (AQP) 1, AQP4, inward rectifying potassium channel 4.1 (Kir4.1) and cytoskeleton features of rat spinal cord astrocytes after cytochalasin D (CytD) intervention. Methods Spinal cord astrocytes isolated from 2~3-day-old rats were cultured till confluency. MTT was used to assess survival rate of astrocytes 2 h, 12 h and 24 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD, respectively. Confocal microscopy was used to observe cytoskeleton features of astrocytes 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml of CytD. The expression of AQP1, AQP4, Kir4.1 mRNA were determined with real-time PCR 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD. Results The survival rate of rat spinal cord astrocytes reduced with the time of co-culture and concentration of CytD (P<0.05). The cytoskeleton of astrocytes was reconstructed. The expression of AQP1, AQP4 and Kir4.1 mRNA increased after co- cultured with 0.05~0.40 μg/ml of CytD. Conclusion The appropriate dosage of CytD may remodel the cytoskeleton and increase the mRNA expression of AQP1, AQP4 and Kir4.1 in spinal cord astrocytes of rats.